Nunzio Turiaco

Evidence for a Role of Mitogen-Activated Protein Kinase 3/Mitogen-Activated Protein Kinase in the Development of Testicular Ischemia-Reperfusion Injury

Abstract Mitogen-activated protein kinase (MAPK) 3/MAPK1 (also known as ERK1/ERK2) plays an important role in the signal transduction pathways. To our knowledge, however, its role in the development of testicular ischemia-reperfusion injury has not yet been investigated. Therefore, we studied the pattern of MAPK3/MAPK1 activation in a experimental model of testicular ischemia-reperfusion injury. We also investigated MAPK8 to understand whether an association exists between these two MAPKs. Adult male Sprague-Dawley rats were subjected to 1 h of testicular ischemia followed by 24 h of reperfusion or to a sham testicular ischemia-reperfusion. Animals were randomized to receive PD98059, which is an inhibitor of MAPK3/MAPK1 (10 mg/kg i.p. administered immediately after detorsion), or its vehicle. The time course of MAPK3/MAPK1, MAPK8, and tumor necrosis factor (TNF; also known as TNF alpha) expression and a histological examination in both the ischemic-reperfused testis and the contralateral one were performed. In both testes, MAPK3/MAPK1 and MAPK8 expression appeared following 10 min of reperfusion and reached their highest activation after 30 min. The MAPK levels slowly decreased, and no significant expression of either kinase was observed following 2 h of reperfusion. Expression of TNF was evident after 1 h of reperfusion and reached its maximum increase after 3 h. PD98059 blunted MAPK3/MAPK1 and MAPK8, reduced TNF expression, and improved the testicular damage caused by ischemia-reperfusion injury in both testes. These data emphasize that MAPK3/MAPK1 has a role in testicular damage and that its blockade might have a future therapeutic role for the management of patients with unilateral testicular torsion.

A psychological preoperative program: effects on anxiety and cooperative behaviors

To evaluate the effectiveness of a psychological preoperative program in reducing preoperative anxiety and in promoting compliance of pediatric participants with surgical procedures.

Polymorphism rs2274911 of GPRC6A as a Novel Risk Factor for Testis Failure

CONTEXT The G protein-coupled receptor GPRC6A is an emerging effector with multiple endocrine roles, including stimulation of T production from the testis. Recently, two men with an inactivating mutation (F464Y) of GPRC6A have been identified, and they showed primary testicular failure and deranged spermatogenesis. Furthermore, one of them also reported cryptorchidism at birth. In addition, a polymorphism (rs2274911, Pro91Ser) in GPRC6A is associated with prostate cancer, a typical androgen-sensitive cancer. OBJECTIVE To study the possible association between rs2274911 polymorphism and male fertility and/or cryptorchidism. Design, Patients, Settings: A total of 611 subjects, including 343 infertile patients, 197 normozoospermic controls, and 71 cryptorchid newborns, were retrospectively selected. METHODS Sequencing analysis for rs2274911 polymorphism and F464Y mutation, and serum levels of FSH, LH, and T were assessed. In vitro functional studies for rs2274911 and F464Y were also performed. RESULTS Homozygous subjects for the risk allele A of rs2274911 had a 4.60-fold increased risk of oligozoospermia and 3.52-fold increased risk of cryptorchidism. A significant trend for increased levels of LH in the GA and AA genotypes, compared with GG homozygotes, was detected in men with azoospermia/cryptozoospermia (P for trend = .027), further supporting an association with primary testicular failure. The mutation F464Y was found in one cryptorchid child (one in 71; 1.41%). Functional studies showed that the A allele of rs2274911 and the F464Y substitution were associated with lower exposition of the receptor on the cell membrane and a reduced downstream phosphorylation of ERK1/2 with respect to wild type. CONCLUSION Our results suggest that GPRC6A inactivation or sub-function contributes to reduced exposure to androgens, leading to cryptorchidism during fetal life and/or low sperm production in adulthood.

Myofibroblasts in adolescent varicocele: an ultrastructural and immunohistochemical study

Abstract Myofibroblasts of the testes play an important role in the morphofunctional integrity of the seminiferous tubule. Previous studies in adults with varicocele have demonstrated an involvement of this cell population that tends to transform into fibroblasts. The aim of the present study was to try and verify the morphological features of myofibroblasts in the adolescent with left idiophatic varicocele. Twenty-two testicular biopsies were obtained from adolescents (aged 13–18 years, mean 15.8) and operated for left idiophatic varicocele. Biopsies were processed for electron microscopy (TEM) and immunofluorescence studies. The latter determined the level of myofibroblasts by using α-smooth muscle isoactin, a specific marker of myofibroblasts and, hence, excluding fibroblasts. TEM observations revealed a normal ultrastructure of myofibroblasts that was similar to that for the controls but an increased presence of extracellular matrix. The immunofluorescence study always demonstrated strong cell positivity to anti α-smooth muscle isoactin as also seen in the controls. This study demonstrates that adolescents with varicocele have well preserved myofibroblasts and do not show any evidence of transformation into fibroblasts, this has already been demonstrated in adult varicocele. These observations could represent an important factor for the understanding of the reversal of growth failure of the testes observed after early treatment.

Intramural Ganglion Structures in Esophageal Atresia: A Morphologic and Immunohistochemical Study

Introduction and Aim. Disorders of esophageal motility causing dysphagia and gastroesophageal reflux are frequent in survivors to esophageal atresia (EA) and distal tracheoesophageal fistula (TEF). The aim of the present study was to investigate the histologic and immunohistochemical features in both esophageal atretic segments to further understand the nature of the motor disorders observed in these patients. Material and Methods. Esophageal specimens from 12 newborns with EA/TEF and 5 newborns dead of unrelated causes were examined. The specimens were fixed in 5% buffered formalin, included in paraffin and cut in 5 micron sections that were stained with hematoxilin and eosin (H and E), and immunohistochemical stainings for Actin, S-100 protein, Neurofilament, Neuron-Specific-Enolase, Chromogranin A and Peripherin were evaluated under the microscope. Results. In controls, the distribution of the neural elements was rather homogenous at both levels of the esophagus. In contrast, the atretic segments showed quantitative and qualitative differences between them with sparser nervous tissue in the distal one in comparison with the proximal one and with controls. Conclusions. These results further support the assumption that histomorphological alterations of the muscular and nervous elements within the esophageal wall might contribute to esophageal dysmotility in patients surviving neonatal operations for EA/TEF.

Our Experience in Transcrotal Orchidopexy in Children Affected by Palpable Undescended Testis

INTRODUCTION Classically, surgical approach for palpable undescended testis (pUDT) consists is an inguinal orchidopexy. In fact, a double incision allows an adequate mobilization of the spermatic cord and an easy dissection of a patent processus vaginalis and also to perform a subdartos pouch. For reduce potential mobility of the inguinal approach, in 1989 Bianchi and Squire proposed a transcrotal orchidopexy, using a high scrotal incision. We report our experiences and retrospectively evaluate the feasibility and postoperative success of the transcrotal approach for treatment of pUDT. PATIENTS AND METHODS From January 2012 to June 2014, 217 patients, affected by pUDT were treated at our Institution, for a total of 231 orchidopexies (203 monolateral pUDT, 14 bilateral pUDT). Patients, in whom, under anesthesia, the testis could be moved to the neck of the scrotum, have been treated with a transcrotal approach using a high scrotal incision. All patients have been clinically followed up at 1 week and at 1, 2, 3, and 6 months, at 1 year, and then annually and using scrotal sonography with color-Doppler at 2, 3, and 6 months. RESULTS A total of 205 pUDT (88.7%) were considered eligible for transcrotal. Eight (3.9%) pUDT, that were first approached transcrotally, have been converted to inguinal approach. At follow-up, two moderate scrotal hematomas and one inguinal hernia were noted. No recurrence or testicular atrophy was showed in transcrotal approached testes, while 2 recurrences out of 26 procedures (7.7%) were displayed after inguinal orchidopexy. CONCLUSIONS In our experience, transcrotal approach is possible in almost 90% of pUDT. No major complication, such as recurrence or testicular atrophy, has been complained. Just 3 out of the 205 cases (1.5%) reported minor complications and 1 of which required a successful day-case procedure. Our data confirm that transcrotal orchidopexy can be considered effective, safe, and with a success rate being equivalent or better to classical inguinal approach.

Neonatal bowel strangulation: Rare presentation of congenital diaphragmatic hernia

We report a case of congenital diaphragmatic hernia (CDH) with perinatal bowel strangulation requiring intestinal resection. Ten hours after birth, the newborn started to be lethargic and developed bilious emesis. X‐ray documented distended loops of bowel with air fluid levels in the abdomen and a gasless, non‐homogeneous opacity of the left hemithorax, a right mediastinal shift and loss of the sharp left hemidiaphram line. On gastrographin enema the left colon was above the adjacent left diaphragm. Emergency surgery was performed at 16 h of age. The entire small bowel appeared reddish and compromised. After 24 h, second‐look laparotomy was performed and only 25 cm of small bowel were viable. The postoperative period was uneventful. Neonatal bowel strangulation in CDH should be taken into account when estimating postnatal morbidity and mortality and, even if CDH treatment is not an emergency procedure, if gastrointestinal symptoms prevail over respiratory symptoms, surgery should be carried out without delay.

Is inguinal orchidopexy still a current procedure in the treatment of intraabdominal testis in the era of laparoscopic surgery

PURPOSE To report our experience in surgical management of nonpalpable intraabdominal testis (NPIT) by inguinal orchidopexy without division of the spermatic vessels. METHODS We reviewed the records of NPIT patients who underwent orchidopexy between 2012 and 2015. All patients were evaluated ultrasonographically. When the testis was not detected ultrasonographically, a laparoscopic exploration was performed. If the testis was found on laparoscopy, surgery was resumed through an inguinal incision. A follow-up was performed at 1week, 1, 3 and 6months. RESULTS Twenty-one NPIT patients were treated, mean age 21.0±11.7months. Ultrasound identified 15 cases of NPIT (71%); diagnostic laparoscopy was performed in 6 (29%). All patients underwent an inguinal orchidopexy. At 1week, four testes were in a high scrotal position. At 6months follow-up, one testis was in a high scrotal position and one retracted up to the external inguinal ring. No atrophy was recorded. CONCLUSIONS Despite several attempts to find a surgical technique without any significant complications, all described procedures failed to meet the target. In our experience, inguinal orchidopexy is a safe, reliable and successful surgical procedure for the management of NPIT. It should be preferred to a technique requiring vascular division, burdened with a higher incidence of atrophy. TYPE OF STUDY Treatment study. LEVELS OF EVIDENCE Case series with no comparison group.

Altered Integrins Expression of Patients Affected by Cryptorchidism

Objectives: The study aimed to investigate the expression of the integrin isoforms α7A and β1A, expressed by myogenic precursor cells, and α7B and β1D, expressed by mature muscle cells in the cremaster of patients affected by an undescended testis. Methods: Fifteen samples of cremaster were obtained from patients undergoing surgery for an undescended testis. Thirty control specimens of cremaster were harvested from patients with congenital hydrocele or inguinal hernia. Immunofluorescent analysis was carried out using anti-α7A, β1A, α7B, and β1D integrin antibodies. Sections were observed using confocal laser scanning microscopy. Results: As compared with controls, a significant loss of a α7B (p = 0.0355) and β1D (p = 0.0069) integrins and a higher expression of α7A (p = 0.0003) and β1A (p = 0.0150) was detected in the cremaster of patients affected by an undescended testis. Conclusions: Our data document a critical alteration of the cytoskeleton of cremasteric smooth muscle cells in patients with an undescended testis. This might explain the altered function in smooth muscle cells in cremaster implied during testicular descent. We therefore speculate that the postnatal splicing of α7A to α7B and of β1A to β1D integrins is delayed. This could account for the common clinical scenario of spontaneous descent of the testes in the first months of life.

Right bochdalek hernia associated with kartagener syndrome: developmental and clinical observations.

We present a novel case of the association of right-sided Bochdalek hernia, a diaphragmatic life-threatening malformation, and Kartagener syndrome, which is characterized by congenital bronchiectasis, chronic sinusitis, and situs inversus. The developmental and clinical findings are discussed. When an association of diaphragmatic hernia with situs viscerum inversus is encountered, physicians should be mindful of the possibility of Kartagener syndrome because this condition could significantly affect the morbidity of the patient.